A new stabilizing agent for the tetramethyl benzidine (TMB) reaction product in the histochemical detection of horseradish peroxidase (HRP)

Olucha, Francisco E.; Martínez‐Garciá, Fernando; López‐García, Carlos

doi:10.1016/0165-0270(85)90025-1

Cited by 296 publications

(123 citation statements)

References 16 publications

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“…It was cryoprotected in 30% sucrose PB, before being frozen and sectioned at a thickness of 80 m on a sliding microtome. An ordered series of every third section was reacted to reveal the location of the HRP using the chromagen tetramethylbenzidine (TMB) (for detailed procedures, see Perkins et al, 2006;Olucha et al, 1985). Briefly, the sections were incubated in a solution containing 0.005% TMB, 0.245% ammonium molybdate, and 0.25% ethanol in 0.1 M PB, pH 6.0.…”

Section: Methodsmentioning

confidence: 99%

Vestibular Signals in Primate Thalamus: Properties and Origins

Hui¹,

May²,

Dickman³

et al. 2007

J. Neurosci.

131

162

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Section: Methodsmentioning

confidence: 99%

Vestibular Signals in Primate Thalamus: Properties and Origins

Hui¹,

May²,

Dickman³

et al. 2007

J. Neurosci.

131

162

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“…These sections were osmicated in 1% 0~0, in PBS at pH 7.4 for 20 min. In the third cat, the sections were processed in 0.1 M phosphate buffer (PB) with ammoniumheptamolybdate as stabilizing agent (Olucha et al, 1985;Ralston et al, 1988). The sections processed according to this latter protocol were soaked in 1% 0~0, in PB (pH 5.5,4OC) overnight (Henry et al, 1985).…”

Section: Methodsmentioning

confidence: 99%

Cervicothalamic tract terminals are enriched in glutamate-like immunoreactivity: an electron microscopic double-labeling study in the cat

Broman

Ottersen

1992

J. Neurosci.

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The distribution of glutamate-like immunoreactivity (Glu-LI) in the thalamic ventral posterolateral nucleus (VPL) of cats was studied with the EM immunogold technique in order to identify nerve terminal populations that may use glutamate as a neurotransmitter. The investigation was focused on cervicothalamic tract (CTT) terminals, which were labeled by WGA-HRP transported anterogradely from injection sites in the lateral cervical nucleus (LCN). The amount of Glu-LI in different profiles was evaluated quantitatively by counting the number of gold particles and then calculating the areal density of gold particles over different profile types. The highest density of gold particles was found over terminals with morphologic characteristics of terminals of cortical origin (RS terminals), a finding that further supports the glutamatergic nature of these terminals suggested by previous studies. Enrichment of Glu-LI was also found in CTT terminals and in non-peroxidase-labeled terminals with the same morphologic characteristics as CTT terminals (RL terminals). The labeling density over these terminals was about twice the average tissue density of gold particles. The labeling density over large VPL neuronal cell bodies was on average 127%, and that over vesicle-containing dendritic appendages and truncs (presynaptic dendrites) about 80%, of the average tissue density of gold particles. Immunogold labeling with antiserum against glutamine (Gln) indicated low levels of Gln-like immunoreactivity in CTT terminals and a high Glu:Gln ratio as compared to astrocytes and the average Glu:Gln ratio in the VPL. The present findings provide further support for a transmitter role of glutamate in terminals of ascending somatosensory afferents to the VPL, including the CTT. Taken together with previous findings of an enrichment of Glu-LI in terminals of the spinocervical tract (Broman et al., 1990), our results suggest that synaptic transmission in the spinocervicothalamic pathway is dependent on the release of glutamate both at the levels of the LCN and the VPL.

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“…Free-floating sections were processed to yield reaction product that is stable and electron dense for ultrastructural analysis (Blok et al 1995). Briefly, Vibratome sections were presoaked in a solution of tetramethylbenzidine and ammonium heptamolybdate (Olucha et al 1985) in 0.1 M phosphate buffer made hypertonic with dextrose. Hydrogen peroxide (0.3%, 1/100 ml) was then added every 5 min for 20 min.…”

Section: Tissue Processing and Electron Microscopymentioning

confidence: 99%

Diverse Synaptic Terminals on Rat Stapedius Motoneurons

Lee

Benson

Brown

2008

JARO

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Stapedius motoneurons (SMN) mediate the contraction of the stapedius muscle, which protects the inner ear from injury and reduces the masking effects of background noise. A variety of inputs to SMNs are known to exist, but their terminal ultrastructure has not been investigated. We characterized the synaptic terminals on retrogradely labeled SMNs found just ventromedial to the facial motor nucleus. About 80% of the terminals contained round synaptic vesicles. One type (Sm Rnd) had small, round vesicles filling the terminal with occasional dense core vesicles and formed an asymmetric synapse. Sm Rnd terminals were small with lengths of apposition to the SMN less than 3 μm. Partial reconstructions from serial sections demonstrated that these terminals formed up to three synapses per terminal. Another terminal type (Lg Rnd) had large, round vesicles and asymmetric synapses. Most Lg Rnd terminals were small but some were extensive, e.g., abutting the SMN for up to 10 μm. One of these terminals formed at least seven synapses. Another terminal type (Pleo) had pleomorphic vesicles and symmetric active zones that, in some cases, were invaginated by spines from the SMN. A fourth uncommon terminal type (Het Rnd) had round vesicles of heterogeneous sizes and asymmetric synapses. A fifth rare terminal type (Cist) had large, round vesicles and an accompanying subsurface cistern in the SMN. These were generally the same kinds of terminals found on other motoneurons, but the high proportion of round vesicle synapses indicate that SMNs receive mostly excitatory inputs.

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A new stabilizing agent for the tetramethyl benzidine (TMB) reaction product in the histochemical detection of horseradish peroxidase (HRP)

Cited by 296 publications

References 16 publications

Vestibular Signals in Primate Thalamus: Properties and Origins

Vestibular Signals in Primate Thalamus: Properties and Origins

Cervicothalamic tract terminals are enriched in glutamate-like immunoreactivity: an electron microscopic double-labeling study in the cat

Diverse Synaptic Terminals on Rat Stapedius Motoneurons

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